PROJECT SUMMARY Quantitative measurements of small molecules are essential to pharmaceutical sciences research. Discovery of the mechanisms of disease pathogenesis for development of new therapeutic strategies requires the quantitative measurements of small molecules in complex biological matrices. The most specific and sensitive method of quantification of these targets is through triple quadrupole mass spectrometry system. Thanks to funds from a share instrument grant (S10RR023461), we purchased a Waters Acquity UPLC with a Thermo triple-stage quadrupole (TSQ) Quantum Ultra mass spectrometry and established our Small Molecular Biomarker Core (SMBC) in 2009. Since then our core has developed sensitive and robust quantitation methods for multiple panels in the analysis of biomarkers and endogenous products, drugs and metabolites, and new drug discovery. The highly sensitive and specific techniques employed have allowed preclinical and clinical researchers to accurately measure multiple small molecule drug and biomarker concentrations in the same sample, maximizing the ability to link basic science with clinical outcomes. We have supported over 40 principal investigators with their ongoing NIH-funded research projects, as well as provided preliminary data to support NIH proposals subsequently selected for funding. SMBC has serviced at least 15 NIH funded grants in the last five years. Our core service has been instrumental to support the funded research, securing new funding, and peer-reviewed publication of research results. Despite its essential role in supporting human health related research, the SMBC core cannot continue to fulfill its mission without replacing the aging instrumentation. The specific aim of this proposal is to request funds from NIH to acquire an integrated system including a Vanquish H UHPLC with a TSQ Altis mass spectrometer from Thermo to replace our 12-year old instrumentation, which will soon cease to be supported by the manufacturers. The new LC-MS/MS system will reside in our SMBC core where resources are available for administering use of the instrument, which include: 1) track record of collaborative research with investigators from all of the health sciences; 2) a well-developed administrative plan for system management, maintenance, and cost recovery; and 3) skilled personnel with substantial experience with mass spectrometric and liquid chromatographic methods. Availability of such instrumentation would allow our NIH-funded user group to pursue innovative research that would otherwise be severely limited due to lack of such state-of-the-art instrumentation.